Kitamura A, Yoshimura Y, Okamoto T
Faculty of Applied Biological Science, Hiroshima University, Higashi-Hiroshima, Japan.
Poult Sci. 2002 Mar;81(3):408-13. doi: 10.1093/ps/81.3.408.
The aim of this study was to determine whether the population of mitotic cells changes in correlation with apoptotic cell population in follicular tissues during atresia of white follicles in hens. Hens were injected with 5-bromo-2'-deoxyuridine (BrdU) 1 h before tissue collection. The small white follicles were classified as healthy follicles and as early or late atretic follicles by histological observation. Mitotic and apoptotic cells were determined by immunocytochemistry for BrdU and terminal deoxytranceferase-mediated dUTP nick end labeling (TUNEL), respectively. The BrdU labeling was observed in some of the granulosa cells and thecal fibroblast-like cells in healthy follicles, whereas the population of the labeled cells was reduced in the granulosa and theca layers of atretic follicles. The image analysis confirmed that the frequency of BrdU-positive cells declined significantly in the granulosa and theca layers of early atretic follicles compared with those of healthy follicles. In contrast, the TUNEL-positive cells were negligible in healthy follicles. However, they were localized in the granulosa and theca layers of early and late atretic follicles, and those in the theca layer were more inside than outside. The frequency of TUNEL-positive cells was significantly increased with the progress of atretic changes. These results suggest that the population of mitotic cells decreases in association with increase of apoptotic cells during the atretic process of white follicles.
本研究的目的是确定在母鸡白色卵泡闭锁过程中,卵泡组织中有丝分裂细胞的数量是否与凋亡细胞数量相关变化。在采集组织前1小时给母鸡注射5-溴-2'-脱氧尿苷(BrdU)。通过组织学观察将小白卵泡分为健康卵泡、早期或晚期闭锁卵泡。分别通过BrdU免疫细胞化学和末端脱氧核苷酸转移酶介导的dUTP缺口末端标记(TUNEL)法测定有丝分裂细胞和凋亡细胞。在健康卵泡的一些颗粒细胞和卵泡膜成纤维样细胞中观察到BrdU标记,而在闭锁卵泡的颗粒层和卵泡膜层中标记细胞的数量减少。图像分析证实,与健康卵泡相比,早期闭锁卵泡的颗粒层和卵泡膜层中BrdU阳性细胞的频率显著下降。相反,健康卵泡中TUNEL阳性细胞可忽略不计。然而,它们定位于早期和晚期闭锁卵泡的颗粒层和卵泡膜层,卵泡膜层中的细胞更多位于内部而非外部。随着闭锁变化的进展,TUNEL阳性细胞的频率显著增加。这些结果表明,在白色卵泡闭锁过程中,有丝分裂细胞数量减少与凋亡细胞增加相关。